• Korean Journal of Veterinary Service
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• v.33 no.4
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• pp.319-326
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• 2010

Bacteroiospermia is a frequently finding in fresh raw and extended porcine semen and can results in detrimental effects on semen quality and longevity. This study aims to evaluate the type of bacterial contaminants in raw and extended porcine semen and the reducing effect of antibiotic test. To investigate bacterial contaminants, out of 387 sample (raw semen 201, extended semen 186) were collected from 6 artifical insemination centers in Gyeongsangnam-do, were inoculated onto blood agar and MacKonkey agar, respectively. Bacterial colonies were selected after culturing for 48 hours, at $37^{\circ}C$, followed by Gram staining, KOH test, oxidase test, catalase test and eventually identified using VITEK System. Total 15 genus and 24 species of bacteria were isolated from these semen samlpes. In raw semen, the most prevalent contaminants were Pseudomonas aeruginosa, Escherichia coli, Proteus mirabilis, Staphylococcus auricularis, Delftia acidovorans, Acinetobacter lowffii, S. aureus and others. And in extended porcine semen, A. lowffii, S. aureus, S. auricularis and other bacteria were identified. Most of them was G(-), which is nonpathogenic bacteria. It seems that bacterial contaminants in fresh raw and extended porcine semen originated from multiple sources at the farms/stud, and were from animal origin and non-animal origins. Whereas, the 7 virus which is known to be detected in porcine semen in 75 cases was not detected. This results showed that removal of bacterial contamination in raw and extended porcine semen is essential and farms were kept for biosecurity and individual hygienes.

• Journal of Veterinary Clinics
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• v.18 no.4
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• pp.345-349
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• 2001

The aim of this study was to investigate the effect of sperm preservation according to the various kinds of commercially available semen extenders and to investigate the effect of sperm preservation according to the various temperature storages of fresh-extended porcine semen. To investigate the effect of sperm preservation according to the various kinds of commercially available semen extenders, porcine semens diluted in 3 semen extenders, Beltsville Thawing Solution(BTS), Androhep and Kiev, were cooled at $8^{\circ}C$ storage temperature with a controlled cooling rate of $2-4^{\circ}C$/h. Motility, progressive motility, normal sperm(%) and sperm morphology were assessed comparatively. In motility and progressive motility, Androhep extenders revealed better result than other extenders. In normal sperm(%) and sperm morphology, 3 semen extenders revealed similar results. To investigate the effect of sperm preservation according to the various temperature storages of fresh-extended porcine semen, porcine semens diluted in BTS extender, were cooled at 3 storage temperatures($8^{\circ}C$, $12^{\circ}C$ and $17^{\circ}C$) with a controlled cooling rate of $2-4^{\circ}C$/h. Motility, progressive motility, normal sperm(%) and sperm morphology were assessed comparatively. In motility and progressive motility, $8^{\circ}C$ treatment group revealed better result than $12^{\circ}C$ and $17^{\circ}C$ treatment groups. In normal sperm(%) and sperm morphology, 3 temperatures of treatment groups revealed similar results.

• Reproductive and Developmental Biology
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• v.39 no.1
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• pp.7-11
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• 2015

Preservation of liquid semen is an important factor for breeding management in swine industry. Oxidative stress of spermatozoa during liquid preservation has a detrimental effect on sperm quality and decreases fertility. Objective of this study was to determine the effect of antioxidant, Quercetin, on capability of porcine liquid semen preservation. Freshly collected porcine semen from boars (n=3), having proven fertility was counted, diluted to $3{\times}10^7/mL$ and divided into 5 different semen extenders. Aliquots of diluted semen with different extenders were subjected to measure the pH, motility, viability and sperm DNA structure status on elapse time after preservation for 10 days. For the first 3 days, semen preserved in all 5 different extenders maintained their initial pH and either gradually decreased or increased thereafter, indicating lipid peroxidation has started. Sperm motility (r=0.52, p=0.01) and viability (r=0.55, p=0.03) had positive correlation with semen pH. Sperm motility was maintained well (p<0.05) in especially 2 extenders containing Tris and antioxidant compared to other extenders, suggesting both Tris and antioxidant worked as pH regulator and had beneficial effects on sperm characteristic during preservation. Sperm DNA structure status accessed by sperm chromatin structure assay on elapsed time after preservation, tended to be higher in semen preserved without antioxidant. Taken together, addition of antioxidant to extender prevents the sperm from oxidative stress during storage in mechanism by which antioxidant slows the lipid peroxidation, and thus reduced the reactive oxygen species in preserved porcine semen resulted in maintaining semen pH, sperm motility and viability for 7~10 days.

• Reproductive and Developmental Biology
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• v.29 no.2
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• pp.133-139
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• 2005

Oxidative stress is one of the major causes of failure in in vitro storage of boar semen. Reactive oxygen species (ROS) are known to be important mediators of such stress. The present study examined the effects of pyruvate and taurine on sperm motility and expression of BAD, Cytochrome c, Caspase-3 and Cox-2 protein in in vitro storage of boar semen, and tested the effect of semen treated with antioxidant with or without hydrogen peroxide on the development of IVM/IVF porcine embryos. Semen samples were transported to the laboratory at $17^{\circ}C$ within 2 hr after collection and were treated with different concentration of pyruvate $(1\~10mM)$ and taurine $(25\~100mM)$ with or without 250uM $H_2O_2$ respectively. The supplementation of pyruvate and taurine increased sperm motility in boar semen during in vitro incubation at $37^{\circ}C$. Expression of apoptosis protein (BAD, cytochrome c, caspase-3 and cox-2) were reduced in the group of boar semen treated with pyruvate and taurine when compared to the other groups. The developmental rates of IVM/IVF porcine embryos fertilized by semen treated with pyruvate and taurine were significantly increased when compared to control (P<0.005). These results indicate that supplementation of pyruvate and taurine as antioxidants in boar semen extender can improve the semen quality and increase in vitro development of porcine IVM/IVF embryos when boar semen treated with antioxidants was used for in vitro fertilization.

• Journal of Animal Science and Technology
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• v.50 no.6
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• pp.793-798
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• 2008

The semen collection process in the porcine is far from being a sterile procedure. Consequently, porcine ejaculates commonly contain bacterial contaminants. The aim of this study is to identify the bacteria in porcine semen and to find the antibiotics resistance of bacteria. Twelve porcine originating from four AI center were used to collect semen. Bacteria were identified by automated instrument for rapid organism identification system and bacterial sensitivities of 8 antibiotics were tested. The Bacterial contaminants of Staphylococcus genus(37.8%), Proteus genus(7.0%), Bacillus genus (6.1%), Pasteulla genus(5.7%), Acinetobacte genus(5.2%), Serratia genus(4.3%) and others(33.9%) were frequently isolated. However, amikacin showed higher antibiotic sensitivity than other antibiotics. General sanitation protocols can contribute partly to inhibit the bacterial contamination, with monitoring boar housing, semen collection areas and the extended semen. But, proper selection of preservative antibiotics by microbial sensitivities can minimize the influence of bacteria.

• Korean Journal of Veterinary Service
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• v.31 no.4
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• pp.547-554
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• 2008

Bacteriospermia is a frequent finding in fresh boar semen and can result in detrimental effects on semen quality and longevity. The objectives of this study was to evaluate types of bacterial contaminants in porcine fresh semen and the reducing effect of antibiotic and density gradient with percoll on the bacterial contaminants. Fresh semen was collected by gloved-hand method into a pre-warmed($37^{\circ}C$) thermostable bottle, and was inoculated onto blood agar and MacConkey agar, respectively. After incubated for 48 hour, 7.5% $CO_2$ at $37^{\circ}C$, bacterial colonies were selected and identified by Gram staining, oxidase test, catalase test and finally identified using API kits and Vitek system. Aerobic culture yielded a variety of bacteria from different genera. The most prevalent contaminant of fresh semen were Leclecia adecarboxylata, Acineobacter banmanni, Staphylococcus epidermidis, Staphylococcus cohni spp urealyticus, Proteus mirabilis. Most of identified bacteria were Gram(-) and non-pathogenic bacteria. It seems that bacterial contaminants in fresh semen were seem originated from multiple sources at the stud/farm, and were from animal and non-animal origins. Gentamicin treatment did not eliminate the bacterial contaminants completely but 3 step-density gradient with percoll completely removed the bacterial contaminants in fresh semen. Therefore, future study is necessary to prove that density gradient method with percoll can eliminate bacteria in fresh semen without significantly affecting sperm viability or function.

• Reproductive and Developmental Biology
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• v.35 no.4
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• pp.479-483
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• 2011

The objective of this study was to determine the effects of E. coli isolated from porcine semen on sperm viability, motility, and semen pH. Semen samples were prepared using commercial extender, $Seminark^{Pro}$ (Noahbio Tech, Korea) that did not contain antibiotics. And 4 different levels of E. coli were artificially innoculated to semen with following concentrations; 4,000 of sperms with 1 of E. coli (T1), 400 with 1 (T2), 40 with 1 (T3), and 4 with 1 (T4). Semen samples were preserved at $17^{\circ}C$ for 5 days in semen storage box until analyzed by flowcytometer. Aliquots were subjected to measure the sperm viability (Live/$Dead^{(R)}$ stain), motility (mitochondrial function), and semen acidity (pH) from day 0 (day of semen collection) to day 5. Sperm motility and viability were significantly decreased (p<0.05) on day 0 (4 hrs after preservation at $17^{\circ}C$) in T3 and T4 compared to control groups and were significantly decreased (p<0.05) in all groups from day 3. Sample pH was acidic in T3 (6.90~6.86) and T4 (6.86~6.65) from day 3 to day 5 (p<0.05). On the other hand, sample pH was maintained 7.0~7.1 in control, T1, and T2 during the experimental period. Sperm motility and viability were significantly decreased from day 0 to day 5 compared to control in samples contaminated with E. coli above a value of 40:1 ($20{\times}10^6$ sperm cells/ml : $5{\times}10^5$ cfu/ml). Even on day 1 in T4 and on day 3 in T3, semen pH was acidic probably due to the acidification of dead spermatozoa. These results suggest that E. coli contamination has a concentration-dependent detrimental effect on extended porcine semen quality.

• Journal of Marine Bioscience and Biotechnology
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• v.6 no.2
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• pp.53-61
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• 2014

This study was investigated the antimicrobial activity of glutaraldehyde cross-linked glucosamine. Glutaraldehyde was used as a cross-linker which specifically combines an amine-group of molecules. To optimize the mixing ratio of glutaraldehyde and glucosamine, mixing ratio was set up 1:1, 2:1, 3:1 and 0.5:1 in molarity, respectively. The optimum mixing ratio of glucosamine and glutaraldehyde was found to be 3:1 using thin layer chromatography based on the production of complex. Glucosamine-glutaraldehyde cross-linked complex (Ggcc) revealed significant antimicrobial activity toward PWG than F1, both microbial strains were isolated from porcine semen as antibiotics resistance bacteria (ARB). These results clearly demonstrate that Ggcc has potential bactericidal activity toward ARB in porcine semen.

• Journal of Embryo Transfer
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• v.24 no.3
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• pp.199-205
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• 2009

This study was carried out to investigate the effective genetic resources preservation system using the frozen boar semen. The porcine oocytes were matured for 44 hours in NCSU-23 medium with or without 10% Porcine Follicle Fluid (PFF), 0.5 ${\mu}g/ml$ porcine FSH, 0.5 ${\mu}g/ml$ equine LH, 1.0 ${\mu}g/ml$ 17 $\beta$-estradiol ($E_2$) and 10 ng/ml Epidermal Growth Factor (EGF) under mineral oil at $38.5^{\circ}C$ in humidified atmosphere of 5% $CO_2$ in air. After 44 h of culture, the oocytes were inseminated with frozen-thawed semen and fresh semen prepared with mTBM medium for 6 h. Later, set of 50 presumptive zygotes were transferred into 4-well dish (500 ${\mu}l$) of IVC medium. for embryos freezing, slow-freezing and vitrification methods were used as a cryopreservation. Differences among treatments were analyzed using General Linear Model Procedure by SAS Package (version 6.12) differences were considered significant when p<0.05. Following IVF and IVC, the rates of cleavage and blastocysts formation were significantly higher (p<0.05) in hormone supplemented group than that of hormone-free group (25.7 vs, 12.1). The development rates to cleavage and blastocysts were significantly higher in PZM-5 group than NCSU-23 group (60.3%, 46.6% vs 27.4%, 11.1%). Further improvement was achieved when PZM-5 was supplemented with FBS. Cleavage rates was significantly higher in fresh semen source group than frozen semen (66.7% vs 43.7%). However in blastocysts rates was similar two groups. Post-thaw survival rates of embryos were 1.2% and 2.2% in slow-frezing and vitrification groups, respectively. The results of our study suggest that it is still possible to improve the culture conditions and boar semen cryopreservation for enhance reproductive technology and animal genetic resources conservation.

• Korean Journal of Veterinary Service
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• v.34 no.2
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• pp.111-116
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• 2011

Artificial insemination (AI) of swine is a very useful reproductive tool and that offers convenience in the Korean swine industry. Since many viruses have been reported to be excreted through boar semen, we investigated the presence of antibodies and antigens against viruses causing reproductive failure in semen of boar in 349 semen samples collected from six Korean AI centers. Viral antigens were detected by polymerase chain reaction (PCR) or reverse transcription-PCR predominantly. The results was as follows. The major reproductive failure causing factor was porcine circovirus type 2 (PCV2), followed by porcine reproductive and respiratory syndrome virus (PRRSV) ($X^2$=166.64, P<0.001). PCV2 and PRRSV, Japanese encephalitis virus (JEV), encephalomyocarditis virus (EMCV) was detected in 73 samples (20.9%), 44 samples (12.6%), 4 samples (1.1%), 3 samples (0.9%), respectively and porcine parvovirus in one sample (0.3%) Classical swine fever virus (CSFV), bovine viral diarrhea virus and Aujeszky's disease virus (ADV) were not detected. Enzyme-linked immunosorbent assay was carried out in 111 serum samples from three AI centers. In most pigs, antibodies response was showed prominently in CSFV (105 sera, 94.6%) ($X^2$=82.580, P<0.001), followed by, in PRRSV (100 sera, 90.1%), PCV2 (92 sera, 90.1%), and PPV (8 sera, 82.9%). ADV antibody was not detected. Thus, the experimental results will be used for the base data, with respect to the state of viral stillbirth in general pig farms, as well as AI centers and breeding farms in Korea.